应用领域： These cells can be used to investigate metabolic and mitogenic signaling of insulin metabolism and its metabolites. This cell line stably expresses the human insulin receptor B (M10051). Special Applications: Potency determination for human insulin and human insulin analogs such as insulin glargine [Lantus, (GlyA21,Arg B31,Arg B32) insulin].
|培养基||The base medium for this cell line is ATCC-formulated F-12K Medium, (ATCC® 30-2004™). To make the complete growth medium, add the following components to the base medium: 56 ml heat inactivated fetal bovine serum FBS and 3.4 ml 50 mg/mL Hygromycin B stock solution.|
|传代方法||Volumes used in this protocol are for 75 cm2 flasks; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes. Remove and discard culture medium. Briefly rinse the cell layer with Ca++/Mg++ free Dulbecco's phosphate-buffered saline (D-PBS) or 0.05% (w/v) Trypsin - 0.53 mM EDTA solution to remove all traces of serum which contains trypsin inhibitor. Add 1.0 to 2.0 mL of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes). Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37°C to facilitate dispersal. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting. Add appropriate aliquots of the cell suspension to new culture vessels. Incubate cultures at 37°C. Subcultivation ratio: A subcultivation ratio of 1:3 to 1:8 is recommended. Medium renewal: Every 2 to 3 days|
|生长条件||Temperature: 37°C Atmosphere:air, 95%; carbon dioxide (CO2)|
|存储条件||liquid nitrogen vapor phase;Freeze medium: complete growth medium supplemented with 5% (v/v) DMSO Storage temperature: liquid nitrogen vapor phase|